Production Of Carboxymethylcellulase Research Articles

Correction: Optimization of Carboxymethyl Cellulase Production by Bacillus sp. NAB37 and Biocatalytic Potential in Saccharification of Acid-Alkali Pretreated Parthenium Hysterophorus

Correction: Optimization of Carboxymethyl Cellulase Production by Bacillus sp. NAB37 and Biocatalytic Potential in Saccharification of Acid-Alkali Pretreated Parthenium Hysterophorus

Optimization of Carboxymethyl Cellulase Production by Bacillus sp. NAB37 and Biocatalytic Potential in Saccharification of Acid-Alkali Pretreated Parthenium Hysterophorus

Optimization of Carboxymethyl Cellulase Production by Bacillus sp. NAB37 and Biocatalytic Potential in Saccharification of Acid-Alkali Pretreated Parthenium Hysterophorus

Design and application of an efficient cellulose-degrading microbial consortium and carboxymethyl cellulase production optimization

Microbial consortia with high cellulase activities can speed up the composting of agricultural wastes with high cellulose contents and promote the beneficial utilization of agricultural wastes. In this paper, rabbit feces and sesame oil cake were used as feedstocks for compost production. Cellulose-degrading microbial strains were isolated from compost samples taken at the different composting stages and screened via Congo red staining and filter paper degradation test. Seven strains, Trichoderma reesei, Escherichia fergusonii, Proteus vulgaris, Aspergillus glaucus, Bacillus mycoides, Corynebacterium glutamicum, and Serratia marcescens, with high activities of carboxymethyl cellulase (CMCase), filter paper cellulase (FPase), and β-glucosidase (β-Gase) were identified and selected for consortium design. Six microbial consortia were designed with these strains. Compared with the other five consortia, consortium VI composed of all seven strains displayed the highest cellulase activities, 141.89, 104.56, and 131.18 U/ml of CMCase, FPase, and β-Gase, respectively. The single factor approach and response surface method were employed to optimize CMCase production of consortium VI. The optimized conditions were: culture time 4.25 days, culture temperature 35.5°C, pH 6.6, and inoculum volume 5% (v/v). Under these optimized conditions, the CMCase activity of consortium VI was up to 170.83 U/ml. Fermentation experiment of rabbit feces was carried out by using the consortium VI cultured under the optimal conditions. It was found that the application effect was better than other treatments, and the fermentation efficiency and nutrient content of the pile were significantly improved. This study provides a basis for the design of microbial consortia for the composting of agricultural wastes with high cellulose contents and provides a support for beneficial utilization of agricultural wastes.

Degradation of 2,6-dicholorophenol by Trichoderma longibraciatum Isolated from an industrial Soil Sample in Dammam, Saudi Arabia

2,6-Dichlorophenol (2,6-DCP) is an aromatic compound with industrial importance in making insecticides, herbicides, and other organic compounds. However, it poses serious health and ecological problems. Microbial degradation of 2,6-DCP has been widely applied due to its effectiveness and eco-friendly characteristics. In this study, Trichoderma longibraciatum was isolated from an industrial soil sample in Dammam, Saudi Arabia using the enrichment method of mineral salt's medium (MSM) amended with 2,6-DCP. Morphological and molecular identification (using the internal transcribed spacer rRNA gene sequencing) of the 2,6-DCP tolerating fungal isolate were charactraized. The fungal isolate has demonstrated a tolerance to 2,6-DCP up to 300 mg/L. Mycelial growth and fungal sporulation were reduced with increasing 2,6-DCP concentrations up to 96 h incubation period. However, after 168 h incubation period, the fungal isolate recorded maximum growth at all the tested 2,6-DCP concentrations up to 150 mg/L. Carboxy methyl cellulase production by tested fungus was decreased by increasing 2,6-DCP concentration up to 75 mg/L. The biodegradation pattern of 2,6-DCP in GM liquid medium using GC–mass analysis as well as the degradation pathway was presented. This study provides a promising fungal isolate that could be used in the bioremediation process for chlorinated phenols in soil.

Medium supplementation and thorough optimization to induce carboxymethyl cellulase production by Trichoderma reesei under solid state fermentation of nettle biomass

In the present study, the production of cellulase by Trichoderma reesei under solid-state fermentation of nettle biomass was promoted through supplementation of the culture media using carbonaceous additives and comprehensive optimization of the cultivation via the Taguchi method. CMCase activities about 5.5–6.1 U/gds were obtained by fermentation of the autoclave-pretreated biomass, among various chemical and physical pretreatments. Then, several additives including Tween 80, betaine, carboxymethyl cellulose, and lactose were individually or in combination added to the culture media to induce the enzyme production. The results proved that such additives could act as either inducers or inhibitors. Furthermore, CMCase activity surprisingly increased to 14.0 U/gds by supplementing the fermentation medium with the optimal mixture of additives including 0.08 mg/gds Tween 80, 0.4 mg/gds betaine, and 0.2 mg/gds carboxymethyl cellulose. Factor screening according to Plackett–Burman design confirmed that the levels of Urea and MgSO4 among basal medium constituents as well as pH of the medium were significantly affected CMCase production. By optimizing the levels of these factors, CMCase activity of 18.8 U/gds was obtained, which was noticeably higher than that of fermentation of the raw nettle. The applied procedure can be promisingly used to convert the nettle biomass into valuable products.

ISOLATION AND MOLECULAR CHARACTERIZATION OF POTENTIAL CELLULOLYTIC BACILLUS SPECIES ISOLATED FROM SOIL SAMPLES

The twenty-two bacterial isolates were screened for cellulolytic activity using Congo red stain on Carboxymethylcellulose (CMC) agar plates inoculated with the isolates. All the isolates were found to hydrolyze Carboxymethylcellulose. Five bacterial isolates (FAY88, FAY103, FAY117. FAY136 and FAY182) were selected according to their higher production of carboxymethyl cellulase (CMCase). A Gram stain test was carried out to identify five isolates as Gram-positive rods, all associated mainly with members of the Bacillus. Their CMCase activities were 1.49, 1.26, 1.21, 1.21 and 1.24 U/ml, respectively. Genetic distances for the five selected strains with high cellulase activity based on the 79 bands obtained from the results of RAPD revealed the similarity according to the morphological characterization agrees with genetic distance. The SDS-PAGE analysis was revealed that, the molecular weight of crude cellulase isolated from Bacillus isolates was found approximately of 54 kDa.

Enhanced Production of Carboxymethylcellulase by Recombinant Escherichia coli Strain from Rice Bran with Shifts in Optimal Conditions of Aeration Rate and Agitation Speed on a Pilot-Scale

The optimal conditions including the aeration rate and agitation speed of bioreactors for the production of carboxymethylcellulase (CMCase) by a recombinant Escherichia coli KACC 91335P, expressing CMCase gene of B. velezensis A-68, were different from those for its cell growth. The enhanced production of CMCase by E. coli KACC 91335P with the conventional multistage process needs at least two bioreactors. Shifts in the optimal conditions of the aeration rate and agitation speed of the bioreactor from the cell growth of E. coli KACC 91335P to those for its production of CMCase were investigated for development of the simple and economic process with the high productivity and low cost. The production of CMCase by E. coli KACC 91335P with shifts in the optimal conditions of the aeration rate and agitation speed from the cell growth to its production of CMCase in a 100 L pilot-scale bioreactor was 1.36 times higher than that with a fixed optimal conditions of the aeration rate and agitation speed for the production of CMCase and it was even 1.54 times higher than that with a fixed optimal conditions of the aeration rate and agitation speed for cell growth. The best time for the shift in the optimal conditions was found to be the mid-log phase of cell growth. Owing to the mixed-growth-associated production of CMCase by E. coli KACC 91335P, shifts in the optimal conditions of the aeration rate and agitation speed of bioreactors from the cell growth to its production of CMCase seemed to result in relatively more cells for the participation in its production of CMCase, which in turn enhanced its production of CMCase. The process with a simple control for shifts in the aeration rate and agitation speed of a bioreactor for the enhanced production of CMCase by E. coli KACC 91335P on the pilot-scale can be directly applied to the industrial-scaled production of cellulase.

Carboxymethyl Cellulase Production by Extremotolerant Bacteria in Low-Cost Media and Application in Enzymatic Saccharification of Stevia Biomass

The aim of this study was to characterize novel cellulase-producing bacteria isolated from desertic environments in Tunisia, and then applied for cellulosic ethanol production. In the first instance, 27 alkali-thermo-halotolerant bacterial strains, belonging to Bacillaceae family, were screened on agar plates containing carboxymethyl cellulose as the sole carbon source. The selected high cellulase producing bacteria were identified as Bacillus mojavensis AG21, Bacillus vallismortis J77, Bacillus sonorensis 7.26 and Bacillus safiensis AJ49. Production of cellulolytic enzymes by selected bacteria was further tested in submerged cultures with Posidonia oceanica leaves, Peganum Harmala biomass, Stevia rebaudiana leaves (SRL), Olive cake, Spent coffee ground and peanut hulls as low-cost cellulosic substrates. Response surface methodology was applied to evaluate the effect of SRL concentration, NaCl concentration, cultivation pH and temperature on carboxymethyl cellulase production and to optimize enzyme yields by selected bacteria. Maximum cellulase production yields of about 1890, 1642, 1516 and 1455 IU/L were recorded for B. mojavensis AG21, B.vallismortis J77, B. sonorensis 7.26 and B. safiensis AJ49, respectively. Production of reducing sugars by hydrolysis of Stevia rebaudiana leaves with bacterial cellulases and commercially fungal cellulases was investigated and compared. The overall reducing sugar yields of B. mojavensis, B.vallismortis, B. sonorensis, B. safiensis, Aspergillus niger and Trichoderma reesei treated- SSR biomasses reached 3620 ± 181, 3040 ± 164, 3656 ± 164, 2654 ± 106, 2646 ± 145 and 2253 ± 123 mg/L respectively. The unique property of cellulase-producing extremotolerant bacteria, proves the potential candidature for current mainstream agricultural cellulosic biomass conversion into fuel ethanol.

Carboxymethyl cellulase production optimization from Glutamicibacter arilaitensis strain ALA4 and its application in lignocellulosic waste biomass saccharification

In this context, carboxymethyl cellulase (CMCase) production from Glutamicibacter arilaitensis strain ALA4 was initially optimized by one factor at a time (OFAT) method using goat dung as proficient feedstock. Two-level full factorial design (25 factorial matrix) using first-order polynomial model revealed the significant (p < 0.05) influence of pH, moisture, and peptone on CMCase activity. Central composite design at N = 20 was further taken into account using a second-order polynomial equation, and thereby liberated maximum CMCase activity of 4925.56 ± 31.61 U/g in the goat dung medium of pH 8.0 and 100% moisture containing 1% (w/w) peptone, which was approximately two fold increment with respect to OFAT method. Furthermore, the partially purified CMCase exhibited stability not only at high pH and temperature but also in the presence of varied metal ions, organic solvents, surfactants, and inhibitors with pronounced residual activities. The enzymatic hydrolysis using partially purified CMCase depicted the maximum liberation of fermentable sugars from alkali pretreated lignocellulosic wastes biomass in the order of paddy straw (13.8 ± 0.15 mg/g) > pomegranate peel (9.1 ± 0.18 mg/g) > sweet lime peel (8.37 ± 0.16 mg/g), with saccharification efficiency of 62.1 ± 0.8, 40.95 ± 0.4, and 37.66 ± 0.4%, respectively after 72 hr of treatment.

Catalytic properties of cellulases and hemicellulases produced by Lichtheimia ramosa: Potential for sugarcane bagasse saccharification

The use of microbial enzyme cocktails for the conversion of polysaccharides of plant origin into fermentable sugars is a global trend. The objectives of this study were to optimize the production of cellulases and hemicellulases by the fungus Lichtheimia ramosa, evaluate the catalytic properties of the produced enzymes, and apply these biocatalysts in the saccharification of sugarcane bagasse. The production of carboxymethylcellulase (CMCase), β-glucosidase, xylanase, and β-xylosidase by L. ramosa were 168.1 ± 2.2, 270.4 ± 8.9, 34 ± 0.8, and 199.2 ± 2.6 U/g of dry substrate, respectively. The optimum pH for the activity of the enzymes ranged between 4.5 and 5.5, and the optimum temperature varied between 55 °C and 65 °C. The enzymes were stable within a wide range of pH, and approximately 95% of their original activity was preserved when incubated for 1 h at 55 °C. The half-lives (t1/2) of CMCase, β-glucosidase, xylanase, and β-xylosidase were 68, 59, 52, and 54 min, respectively, when incubated at 60 °C. The enzymes were stable in solutions containing ethanol (10%), and the kinetic parameters (Km and Vmax) demonstrated that β-glucosidase from L. ramosa was competitively inhibited by glucose. The enzyme extract produced by L. ramosa was used for the saccharification of sugarcane bagasse pretreated with glycerol and the highest yield of glucose (10.66%) was obtained at 24 h of hydrolysis. The characteristics of the enzymes, combined with the efficiency in hydrolyzing sugarcane bagasse, allow for the application of this enzyme extract to processes of cellulose saccharification for the production of second-generation ethanol.

Molecular Characterization of Trichoderma asperellum and Lignocellulolytic Activity on Barley Straw Treated with Silver Nanoparticles

Silver nanoparticles (AgNPs) have been applied as an antifungal agent, which results in AgNPs contamination of agricultural waste that interferes with the lignocellulosic enzymes produced by fungi. Therefore, this study examined the production of carboxymethylcellulase (CMCase) and manganese-dependent lignin peroxidase (MnPase), using barley straw treated with AgNPs. Trichoderma asperellum growth was not inhibited at 25 ppm AgNPs, while negligible growth inhibition was observed at 50 ppm AgNPs, which was not observed with Aspergillus terreus and Curvularia lunata. T. asperellum was the highest producer of CMCase and MnPase using barley straw with or without 25 ppm AgNPs versus A. terreus or C. lunata. AgNPs addition to barley straw before T. asperellum inoculation played a role in repressing enzyme activities (CMCase 156.33 U/mL and MnPase 1.28 U/mL); however, addition of AgNPs (50 ppm) after 10 days of incubation showed the highest activity (CMCase 160.67 and 1.35 U/mL MnPase). Optimum temperature for enzyme production by T. asperellum using untreated and treated barley straw was 35 °C and 30 °C, respectively. Enzyme activities increased with increasing polyoxyethylenesorbitan monooleate surfactant concentrations up to 0.25 mL/g substrate without AgNPs, whereas the activity decreased with AgNPs (25 ppm). The exception to this observed trend was at low concentrations of the surfactant (0.10 mL/g substrate).

Cellulase Production by Trichosporon laibachii

The ability of twenty-three yeast strains isolated from decayed wood and the fungus garden of attine ants to produce hemi- and cellulolytic hydrolases in a chemically defined medium was screened. Xylanase, β-glucosidase and cellulase activities were found in yeasts strains in the genera Cryptococcus , Trichosporon , Debaryomyces and Pichia . Among the isolated microorganisms, two strains of Trichosporon laibachii MG270406-1A14 strain showed higher cellulase titers. Several biochemical and physiological parameters were investigated for optimum cellulase production under submerged fermentation. Enzyme induction was also examined using different carbon and nitrogen sources. Cellobiose and ammonium sulphate were found best carbon (C) and nitrogen (N) sources to enhance carboxymethylcellulase production. A 2 2 - factorial design with center points was developed to optimize enzyme production, and data analysed by the response surface method. Maximal enzyme titres (0.3 U mL -1 ) occurred at initial pH 6.0, C/N ratio 9.0 and 5 days. DOI: http://dx.doi.org/10.17807/orbital.v9i4.1024

Comparison of optimal conditions for mass production of carboxymethylcellulase by Escherichia coli JM109/A-68 with other recombinants in pilot-scale bioreactor

The optimal conditions for mass production of carboxymethylcellulase (CMCase) by E. coli JM109/A-68 were investigated and compared with other E. coli JM109 recombinants producing CMCase. The optimal agitation speed and aeration rate for cell growth of E. coli JM109/A- 68 were 500 rpm and 0.50 vvm in a 7 L bioreactor, whereas those for production of CMCase were 416 rpm and 0.95 vvm. The optimal vessel pressures for cell growth as well as production of CMCase in a 100 L bioreactor were 0.04 MPa. The maximal production of CMCase by E. coli JM109/A-68 under the optimized conditions in a 100 L bioreactor was 11.0 times higher than its wild type, B. velezensis A-68. Optimal conditions for mass production of CMCase by recombinants were different from those for wild strains. The higher production of CMCase by E. coli JM109/A-68 and other recombinant of E. coli seemed to result from its higher cell growth under the optimal conditions for dissolved oxygen and its mixed-growth associated production pattern compared to the growthassociated production of B. velezensis A-68.

Carboxymethyl cellulase production optimization from newly isolated thermophilic Bacillus subtilis K-18 for saccharification using response surface methodology

In this study, a novel thermophilic strain was isolated from soil and used for cellulase production in submerged fermentation using potato peel as sole carbon source. The bacterium was identified by 16S rRNA gene sequencing technology. Central composite design was applied for enhanced production using substrate concentration, inoculum size, yeast extract and pH as dependent variables. Highest enzyme titer of 3.50 ± 0.11 IU/ml was obtained at 2% substrate concentration, 2% inoculum size, 1% yeast extract, pH 5.0, incubation temperature of 50 °C for 24 h of fermentation period. The crude enzyme was characterized having optimum pH and temperature of 7.0 and 50 °C, respectively. The efficiency of enzyme was checked by enzymatic hydrolysis of acid/alkali treated pine needles which revealed that 54.389% saccharification was observed in acid treated pine needles. These results indicated that the cellulase produced by the Bacillus subtilis K-18 (KX881940) could be effectively used for industrial processes particularly for bioethanol production.

Degradation of bamboo-shoot shell powder by a fungal consortium: Changes in chemical composition and physical structure

Co-culturing microorganism to degrade lignocellulose has been adapted by various researchers globally. The decomposition of bamboo-shoot shell powder (BSSP) by a two-fungus (Pleurotus ostreatus and Aspergillus niger) together was examined. During the degradation, there was a rise in the activities of enzymes carboxymethylcellulase (catalyzing cellulose degradation by the hydrolysis of β-1, 4 glycosidic bonds in cellulosic materials) and laccase (phenol oxidase catalyzing lignin degradation) responsible for metabolism of lignocellulosic material resulting into decrease in acid detergent fiber and lignin contents of BSPP. In addition, the physical properties of the shell fiber were changed from smooth and flexible to rough and crisp. The products of the decomposing fiber were complex, and most of them contained methyl or ethyl groups. The fungi decomposed native silicon compounds simultaneously during biodegradation. The net result of decomposition was a decrease in biomass as well as production of carboxymethylcellulase and laccase, two biotechnologically relevant extracellular enzymes with multiple industrial uses.

Construction of a recombinant Escherichia coli JM109/A-68 for production of carboxymethylcellulase and comparison of its production with its wild type, Bacillus velezensis A-68 in a pilot-scale bioreactor

A gene encoding carboxymethylcellulase (CMCase) of Bacillus velezensis A-68 had been cloned in Escherichia coli JM109. Based on productivity and economic aspect, rice bran and ammonium chloride were chosen to be optimal carbon and nitrogen sources for production of CMCase by E. coli JM109/A-68. The optimal conditions for rice bran, ammonium chloride, and initial pH of medium for production of CMCase were established by the response surface methodology (RSM). The concentrations of four salts in the medium, K2HPO4, NaCl, MgSO4·7H2O, and (NH4)2SO4, for production of CMCase also were optimized. The optimal temperatures for cell growth and production of CMCase were 37°C. The maximal production of CMCase by E. coli JM109/A-68 was 880.2 U/mL, which was 10.5 time higher than its wild type, B. velezensis A-68. The production of CMCase by E. coli JM109/A-68 was compared with that by B. velezensis A-68 in a 100 L pilot-scale bioreactor under the optimized conditions. The production of CMCase by E. coli JM109/A-68 was found to be the mixed-growth associated unlike the growthassociated production of CMCase by B. velezensis A-68.

Production, purification, and characterization of carboxymethyl cellulase from novel strain Bacillus megaterium

In the present investigation a strain of Bacillus megaterium BM05 was evaluated for cellulase production in submerged fermentation using wheat straw as a substrate. Various nutritional and cultural parameters were optimized to obtain maximum production of carboxymethyl cellulase. The strain produced best titer of cellulase at initial medium pH of 5, inoculum size of 1% using 2% wheat straw with 0.8% NaCl in medium for 48 h of fermentation period. Further supplementation of CMC as an additional carbon source and yeast extract as nitrogen sources favored enzyme production. Organic acids found to be inhibitory effect on secretion of cellulase enzyme by B. megaterium BM05. The enzyme exhibited molecular weight of 53.55 kDa as revealed by SDS‐PAGE. The enzyme had optimum pH and temperature of 6.5°C and 50°C with stability range of 6.0–8.0 and 30°C–40°C for 140 min, respectively. The enzyme had half life of 38.9 min at 80°C. Metal profile of the enzyme indicated that K+ (125%) and Na+ (118%) were the activators while Pb+2 (15%) and Hg+2 (13%) were the inhibitors. The endoglucanase had Km and Vmax value of 1.73 and 0.95 U/mL using CMC‐Na as a substrate, respectively. Turnover number (Kcat) and catalytic efficiency (Kcat/Km) of purified enzyme was 1556.11 min and 1.3 × 106 M/s, respectively. © 2016 American Institute of Chemical Engineers Environ Prog, 35: 1741–1749, 2016

Novel Bacillus subtilis IND19 cell factory for the simultaneous production of carboxy methyl cellulase and protease using cow dung substrate in solid-substrate fermentation.

BackgroundHydrolytic enzymes, such as cellulases and proteases, have various applications, including bioethanol production, extraction of fruit and vegetable juice, detergent formulation, and leather processing. Solid-substrate fermentation has been an emerging method to utilize low-cost agricultural residues for the production of these enzymes. Although the production of carboxy methyl cellulase (CMCase) and protease in solid state fermentation (SSF) have been studied extensively, research investigating multienzyme production in a single fermentation process is limited. The production of multienzymes from a single fermentation system could reduce the overall production cost of enzymes. In order to achieve enhanced production of enzymes, the response surface methodology (RSM) was applied.ResultsBacillus subtilis IND19 utilized cow dung substrates for the production of CMCase and protease. A central composite design and a RSM were used to determine the optimal concentrations of peptone, NaH2PO4, and medium pH. Maximum productions of CMCase and protease were observed at 0.9 % peptone, 0.78 % NaH2PO4, and medium pH of 8.41, and 1 % peptone, 0.72 % NaH2PO4, and medium pH of 8.11, respectively. Under the optimized conditions, the experimental yield of CMCase and protease reached 473.01 and 4643 U/g, which were notably close to the predicted response (485.05 and 4710 U/g). These findings corresponded to an overall increase of 2.1- and 2.5-fold in CMCase and protease productions, respectively.ConclusionsUtilization of cow dung for the production of enzymes is critical to producing multienzymes in a single fermentation step. Cow dung is available in large quantity throughout the year. This report is the first to describe simultaneous production of CMCase and protease using cow dung. This substrate could be directly used as the culture medium without any pretreatment for the production of these enzymes at an industrial scale.

Optimization of Conditions and Production of Carboxy Methyl Cellulase by Bacteria Isolated from Higher Termite Soil

The current work deals with the studies of isolation and preliminary characterization of the bacteria isolated from higher termite soil. Termites play an important role in the turnover and mineralization of complex biopolymers, such as wood and other cellulose and hemicelluloses containing materials. Lignocellulose is the most predominant component of the woody and dead plant materials, as well as it is the most abundant biomass on earth, especially in terrestrial ecosystems. Thus the work focuses on using lingo cellulosic waste efficiently for production of Carboxy Methyl Cellulase (CMC). As CMCase have higher hydrolyzing capacity than the other two cellulases, cellobiase and cellobiohydralases it has wide application in industry such as bread and baking industry, laundry etc. Thus the work focuses on identification of efficient bacterial species in this work 6 bacterial species were isolated among which I4 produced higher CMCase enzyme. After the morphological, biochemical and identification test, the I4 isolate was found to be Bacillus sp.

Carboxymethyl cellulase production from a Paenibacillus sp.

Cellulases are industrially important enzymes with a potential to convert cellulose into fermentable sugars. Novel bacterial isolate Paenibacillus sp. CKS1 was tested for cellulase activity and the optimal conditions for carboxymethyl cellulase (CMCase) production were determined. Maximum CMCase activity was obtained in the third passage of the bacterial culture after 3 days of incubation at 30?C. Cellobiose and yeast extract was the optimal source of carbon and nitrogen for induction of CMCase activity. In addition, with initial pH 7 of the medium and 40 ml of working volume in 500 ml culture flasks with shaking at 150 rpm, the maximum CMCase activity in a crude culture supernatant reached value of 0.532?0.006 U/ml. For crude CMCase, optimal temperature was 50?C and optimal pH 4.8, respectively. HPLC analysis confirmed the bacterium is capable to hydrolise CMC to glucose and other soluble sugars.